Relative quantification of CYP1A gene expression in whitefish (Coregonus lavaretus) exposed to benzo[a]pyrene

• Autorzy: Brzuzan P. , Jurczyk Ł. , Łuczyński M. K. , Góra M.
• Języki publikacji: EN

Abstrakty

EN

The expression of CYP1A (cytochrome P4501A) can be induced by a number of aromatic compounds in teleost fishes. We developed a real-time PCR assay for measuring relative quantities (RQ) of CYP1A mRNA in whitefish (Coregonus lavaretus). To test for the usefulness of the assay we performed a treatment study, using benzo[a]pyrene (B[a]P) a model CYP1A inducer. Primers for the CYP1A gene were adapted from the literature, whereas those for [beta]-actin (endogenous control) were designed from a region that was found to be conserved among salmonid [beta]-actin genes. A group of hatchery raised whitefish, with an average body mass of 15 g and total length of 12 cm were given an intraperitoneal injection (10 mg/kg) of B[a]P in corn oil (2 mg B[a]P/ml corn oil) or corn oil alone (Control). After 48 h, whitefish liver, head kidney and brains were collected for mRNA isolation and analysis. In all three tissues sampled, CYP1A mRNA was affected by treatment with B[a]P. Head kidney tissue showed the greatest induction potential (RQ=11.00) from base levels (RQ=1.00), followed by liver (RQ=9.45), and brain (RQ=3.76). These results demonstrated that CYP1A was highly inducible by B[a]P in whitefish head kidney and liver, and to some extent, in brain tissue. The approach presented here has the advantage of providing rapid and accurate measures of CYP1A induction in various tissues of fish responding to PAH contaminant exposure.

Słowa kluczowe

EN

benzo(a)pyrene; Coregonus lavaretus; CYP1A gene expression; RealTime PCR; relative quantification

PL

ekspresja genu CYP1A; sieja; benzo(a)piren; real-time PCR

• Wydawca: University of Warmia and Mazury in Olsztyn, Poland
• Czasopismo: Environmental Biotechnology
• Rocznik: 2005
• Tom: Vol. 1, No. 1
• Strony: 11--15
• Opis fizyczny: Bibliogr. 19 poz., tab., wykr.

Twórcy

autor

Brzuzan P.

• Department of Environmental Biotechnology, Faculty of Environmental Sciences and Fisheries, University of Warmia and Mazury in Olsztyn, Słoneczna 45G, 10-709 Olsztyn, Poland, Phone: (+48) (89) 523 4187, Fax: (+48) (89) 523 4131

autor

Jurczyk Ł.

• Department of Environmental Biotechnology, Faculty of Environmental Sciences and Fisheries, University of Warmia and Mazury in Olsztyn, Słoneczna 45G, 10-709 Olsztyn, Poland

autor

Łuczyński M. K.

• Department of Chemistry, University of Warmia and Mazury in Olsztyn, Pl. Łódzki 4, 10-957 Olsztyn, Poland

autor

Góra M.

• Department of Organic Chemistry, Jagiellonian University, Ingardena 3, 30-060 Kraków, Poland

Bibliografia

• Arinc, A., A. Sen, A. Bozcaarmutlu. 2000. Cytochrome P4501A and associated mixed function oxidase induction in fish as a biomarker for toxic carcinogenic pollutants in the aquatic environment. Pure and Applied Chemistry 72: 985-994.
• Burczynski, M. E., T. M. Penning. 2000. Genotoxic polycyclic aromatic hydrocarbon ortho-quinones generated by aldo-keto reductases induce CYP1A1 via nuclear translocation of the aryl hydrocarbon receptor. Cancer Research 60: 908-915.
• Cao, Z., J. Hong, R. E. Peterson, J. M. Aiken. 2000. Characterization of CYP1A1 and CYP1A3 gene expression in rainbow trout (Oncorhynchus mykiss). Aquatic Toxicology 49: 101-109.
• Campbell, P.M., R. H. Devlin. 1996. Expression of CYP1A1 in livers and gonads of Pacific salmon: quantitation of mRNA levels by RT-cPCR. Aquatic Toxicology 34: 47-69.
• Hahn, M. E., R. R. Merson, S. I. Karchner. 2005. Xenobiotic receptors in fish: Structural and functional diversity and evolutionary insights. In: Biochemical and Molecular Biology of Fishes Volume 6 Environmental Toxicology. (ed. T. W. Moon, T. P. Mommsen), pp. 191-228. Elsevier, New York.
• Higuchi, R., C. Fockler, G. Dollinger, R. Watson. 1993. Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Biotechnology 11: 1026-1030.
• Incardona, J.P., T. K. Collier, N. L. Scholz. 2004. Defects in cardiac function precede morphological abnormalities in fish embryos exposed to polycyclic aromatic hydrocarbons. Toxicology and Applied Pharmacology 196: 191-205.
• Larkin, P., I. Knoebl, N. D. Denslow. 2003. Differential gene expression analysis in fish exposed to endocrine disrupting chemicals. Comparative Biochemistry and Physiology B 136: 149-161.
• Livak, K. J., T. D. Schmittgen. 2001. Analysis of relative gene expression data using Real-Time Quantitative PCR and the 2-C T method. Methods 25: 402-408.
• Mellanen, P., M. Soimasuo, B. Holmbom, A. Oikari, R. Santti. 1999. Expression of the vitellogenin gene in the liver of juvenile whitefish (Coregonus lavaretus L. s.l.) exposed to effluents from pulp and paper mills. Ecotoxicology and Environmental Safety 43: 133-137.
• Moore, M. J., I. V. Mitrofanov, S. S. Valentini, V. V. Volkov, A. V. Kurbskiy, E. N. Zhimbey, E. B. Eglinton, J. J. Stegeman. 2003. Cytochrome P4501A expression, chemical contaminants and histopathology in roach, goby and sturgeon and chemical contaminants in sediments from the Caspian Sea, Lake Balkhash and the Ily River Delta, Kazakhstan. Marine Pollution Bulletin 46: 107-119.
• Murchelano, R. A., R. E. Wolke. 1991. Neoplasms and nonneoplastic liver lesions in winter flounder Pseudopleuronectes americanus, from Boston Harbor, Massachusetts. Environmental Health Perspectives 90: 17-26.
• Myers, M. S., C. M. Stehr, O. P. Olson, L. L. Johnson, B. B. McCain, S-L. Chan, U. Varanasi. 1994. Relationships between toxicopathic hepatic lesions and exposure to chemical contaminants in English sole (Pleuronectes vetulus), starry flounder (Platichthys stellatus), and white croaker (Genyonemus lineatus) from selected marine sites on the Pacific coast, USA. Environmental Health Perspectives 102: 200-215.
• Prasch, A. L., H. Teraoka, S. A. Carney, W. Dong, T. Hiraga, J. J. Stegeman, W. Heideman, R. E. Peterson. 2003. Aryl hydrocarbon receptor 2 mediates 2,3,7,8-tetrachlorodibenzo-p-dioxin developmental toxicity in zebrafish. Toxicological Sciences 76: 138-150.
• Rees, C. B., W. Li. 2004. Development and application of a real-time quantitative PCR assay for determining CYP1A transcripts in three genera of salmonids. Aquatic Toxicology 66: 357-368.
• Rees, C. B., S. D. McCormick, J. P. Vanden Heuvel, W. Li. 2003. Quantitative PCR analysis of CYP1A induction in Atlantic salmon (Salmo salar). Aquatic Toxicology 62: 67-78.
• Soimasuo, R., A.E. Karels, H. Lepanen, R. Santti, A. Oikari. 1998. Biomarker responses in whitefish (Coregonus lavaretus L. s.l.) experimentally exposed in a large lake receiving effluents from pulp and paper industry. Archives of Environmental Contamination and Toxicology 34: 69-80.
• Stegeman, J.J. 1995. Molecular aspects of oxidative drug metabolizing enzymes: Their significance in environmental toxicology, chemical carcinogenesis and health (ed. E. Arin˜, J. B. Schenkman, E. Hodgson), pp. 135-158. Springer-Verlag, NATO ASI, Heidelberg.
• Teraoka, H., W. Dong, Y. Tsujimoto, H. Iwasa, D. Endoh, N. Ueno, J. J. Stegeman, R. E. Peterson, T. Hiraga. 2003. Induction of cytochrome P4501A is required for circulation failure and edema by 2,3,7,8-tetrachlorodibenzo-p-dioxin in zebrafish. Biochemical and Biophysical Research Communications 304: 223-228.